Effects of myogenic induction, differentiation and transplantation of canine umbilical cord blood stem cells on cell-cell junction
10.3969/j.issn.1673.8225.2009.36.021
- VernacularTitle:犬脐血干细胞肌源性诱导分化移植对细胞间连接的影响
- Author:
Jun WAN
;
Ju MEI
;
Jinben MA
;
Nan MA
;
Genfa SHAN
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(36):7108-7112
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Umbilical cord blood-mesenchymal stem cells (UCB-MSCs) following differentiation into cardiomyocytes were transplanted into ischemic myocardium. The transplanted cells can build connection with host cells and repair the infarct myocardium. OBJECTIVE: To detect the cell-cell junction after transplantation of the cardiac-like cell derived from the canine umbilical cord blood stem cells. DESIGN, TIME AND SETTING: A randomized controlled animal study was performed from July 2006 to October 2007 at the Animal Experimental Center of Xinhua Hospital Affiliated to School of Medicine, Shanghai Jiao Tong University. MATERIALS: A total of 2 full-term pregnant canines were used for isolation of UCB-MSCs. A total of 36 adult mongrel canines were divided into cell transplantation group and model control group (n=18) according to the rule of random digits table. METHODS: The MSCs at passage 4 were transfected by Laz-Z. After 3-day culture, MSCs were induced by 10 μmol/L 5-azacytidine (5-aza). The canine models of myocardium infarction were established following 3 weeks of culture. 2 mL (1 ×107)MSCs were transplanted into dogs with acute myocardium infarction by coronary artery infusion and local injection in cell transplantation group. An equal volume of saline was used in the model control group. The specimens were harvested and detected at 2, 4 and 8 weeks, respectively. Cell junction was determined using immunohistochemistry. MAIN OUTCOME MEASURES: The following parameters were measured: gene trensfection, myogenic induction and differentiation results of UCB-MSCs; junction of transplanted cells and host cardiomyocytes. RESULTS: Following 72 hours of transfaction, mass of cells expressed LacZ gene, synthetized galactosidase, and stained blue using X-gal staining. Following 3 weeks of 5-aza induction, the antigen a-Actin, Desmin and Connexin43 were all been positively expressed, but before induction they were all negative. From the myocardial section of 8 weeks after transplantation, the junction was formed between the transplanted cells and the host myocardium as formed between the transplanted cells. In the junction, green-fluorescence positive expression of cadherin and connexin43 could be seen. However, in the model control group, only cadherin and connexin43 expressed positively, but the transplanted UCB-MSCs with red fluorescence could not been observed. CONCLUSION: The UCB-MSCs is able to differentiate into cardiac-like cell in vitro and form cell-cell junction in vivo to communicate with surrounding cells.
